Abstract
Digitalis purpurea L. is a cardenolide-producing medicinal and ornamental plant. Cardenolides, like digoxin, are commonly used to treat congestive heart failure, cardiac arrhythmia, and atrial fibrillation. More recently, its cytotoxic activity on several types of cancer and antiviral effect have been confirmed. As chemical synthesis is not viable, D. purpurea plants are one of the major sources of cardenolides for the pharmaceutical industry. However, cardenolide content is highly variable under natural conditions. Therefore, D. purpurea in vitro culture has been a focus of research since the second half of the twentieth century. This chapter is a compendium of these reports with emphasis on the effects of morphogenesis, culture conditions, and in vitro culture medium composition on cardenolide content. Besides, some studies on the genetic stability of Digitalis plants are summarized. Finally, we describe the biotechnological approaches reported so far to obtain a higher yield of cardenolides in vitro, such as elicitation and metabolic engineering, both recognized as promising strategies.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
Abbreviations
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- 2iP:
-
N6-[2-Isopentenyl] adenine
- ABA:
-
Abscisic acid
- BA:
-
Benzyladenine
- BTOA:
-
2-Benzothiazole-oxyacetic acid
- DW:
-
Dry weight
- FW:
-
Fresh weight
- GA3:
-
Gibberellic acid
- IAA:
-
Indoleacetic acid
- LED:
-
Light-emitting diode
- LS medium:
-
Linsmaier and Skoog medium
- MS medium:
-
Murashige and Skoog medium
- NAA:
-
Naphthaleneacetic acid
- PEG:
-
Polyethylene glycol
- RAPD:
-
Random amplified polymorphic DNA
- RuBisCO:
-
Ribulose-1,5-bisphosphate carboxylase/oxygenase
- TDZ:
-
Thidiazuron
- TIS:
-
Temporary immersion system
References
David B, Wolfender JL, Dias DA (2015) The pharmaceutical industry and natural products: historical status and new trends. Phytochem Rev 14:299–315. https://doi.org/10.1007/s11101-014-9367-z
El-Seedi HR, Khalifa SAM, Taher EA et al (2019) Cardenolides: insights from chemical structure and pharmacological utility. Pharmacol Res 141:123–175. https://doi.org/10.1016/j.phrs.2018.12.015
Kreis W (2017) The foxgloves (Digitalis) revisited. Planta Med 83:962–976. https://doi.org/10.1055/s-0043-111240
WHO (2018) World Health Statistics 2018: monitoring health for the SDGs, sustainable development goals. World Health Organization, Geneva
Heron M, Anderson R (2016) Changes in the leading cause of death: recent patterns in heart disease and cancer mortality. NCHS data brief, no 254. National Center for Health Statistics, Hyattsville, pp 1–8
Sales E, Frieder M, Nebauer SG et al (2011) Wild crop relatives: genomic and breeding resources. Springer, Berlin/Heidelberg
Verma SK, Das AK, Cingoz GS, Gurel E (2016) In vitro culture of Digitalis L. (Foxglove) and the production of cardenolides: an up-to-date review. Ind Crop Prod 94:20–51. https://doi.org/10.1016/j.indcrop.2016.08.031
Rietbrock N, Woodcock BG (1984) Trends in pharmacological sciences in 1784. Trends Pharmacol Sci 5:446–448. https://doi.org/10.1016/0165-6147(84)90500-5
Staba EJ (1962) Production of cardiac glycosides by plant tissue cultures I: nutritional requirements in tissue cultures of Digitalis lanata and Digitalis purpurea. J Pharm Sci 51:249–254. https://doi.org/10.1002/jps.2600510314
Büchner SA, Staba EJ (1964) Preliminary chemical examination of digitalis tissue cultures for cardenolides. J Pharm Pharmacol 16:733–737. https://doi.org/10.1111/j.2042-7158.1964.tb07397.x
Kaul B, Wells P, Staba EJ (1967) Production of cardio-active substances by plant tissue cultures and their screening for cardiovascular activity. J Pharm Pharmacol 19:760–766. https://doi.org/10.1111/j.2042-7158.1967.tb08028.x
Furuya T, Kawaguchi K, Hirotani M (1973) Biotransformation of progesterone by suspension cultures of Digitalis purpurea cultured cells. Phytochemistry 12:1621–1626. https://doi.org/10.1016/0031-9422(73)80379-6
Hirotani M, Furuya T (1975) Metabolism of 5β-pregnane-3,20-dione and 3β-hydroxi-5β-pregnan-20-one by Digitalis suspension cultures. Phytochemistry 14:2601–2606. https://doi.org/10.1016/0031-9422(75)85233-2
Hirotani M, Furuya T (1980) Biotransformation of digitoxigenin by cell suspension cultures of Digitalis purpurea. Phytochemistry 19:531–534. https://doi.org/10.1016/0031-9422(80)87008-7
Yoshikawa T, Furuya T (1979) Purification and properties of sterol: UDPG glucosyltransferase in cell culture of Digitalis purpurea. Phytochemistry 18:239–241. https://doi.org/10.1016/0031-9422(79)80061-8
Corduan G, Spix C (1975) Haploid callus and regeneration of plants from anthers of Digitalis purpurea L. Planta 124:1–11. https://doi.org/10.1007/BF00390062
Nitsch JP, Nitsch C (1969) Haploid plants from pollen grains. Science 163:85–87. https://doi.org/10.1126/science.163.3862.85
Hagimori M, Matsumoto T, Kisaki T (1980) Studies on the production of Digitalis cardenolides by plant tissue culture I. Determination of digitoxin and digoxin contents in first and second passage calli and organ redifferentiating calli of several Digitalis species by radioimmunoassay. Plant Cell Physiol 21:1391–1404. https://doi.org/10.1093/pcp/21.8.1391
Hagimori M, Matsumoto T, Obi Y (1982) Studies on the production of Digitalis cardenolides by plant tissue culture: II. Effect of light and plant growth substances on digitoxin formation by undifferentiated cells and shoot-forming cultures of Digitalis purpurea L. grown in liquid media. Plant Physiol 69:653–656. https://doi.org/10.1104/pp.69.3.653
Hagimori M, Matsumoto T, Obi Y (1982) Studies on the production of Digitalis cardenolides by plant tissue culture III. Effects of nutrients on digitoxin formation by shoot-forming cultures of Digitalis purpurea L. grown in liquid media. Plant Cell Physiol 23:1205–1211. https://doi.org/10.1093/oxfordjournals.pcp.a076462
Hagimori M, Matsumoto T, Mikami Y (1984) Photoautotrophic culture of undifferentiated cells and shoot-forming cultures of Digitalis purpurea L. Plant Cell Physiol 25:1099–1102. https://doi.org/10.1093/oxfordjournals.pcp.a076797
Hagimori M, Mikami Y, Matsumoto T (1984) Jar Fermenter culture of shoot-forming cultures of Digitalis purpurea L. using a revised medium. Agric Biolog Chem 48:965–970. https://doi.org/10.1271/bbb1961.48.965
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Matsumoto M, Koga S, Shoyama Y, Nishioka I (1987) Phenolic glycoside composition of leaves and callus cultures of Digitalis purpurea. Phytochemistry 26:3225–3227. https://doi.org/10.1016/S0031-9422(00)82474-7
Čellárová E, Hončariv R (1991) The influence of n-[2-isopentenyl] adenine on shoot differentiation in Digitalis purpurea L. tissue cultures. Acta Biotechnol 11:331–334. https://doi.org/10.1002/abio.370110408
Gärtner DE, Seitz HU (1993) Enzyme activities in cardenolide-accumulating, mixotrophic shoot cultures of Digitalis purpurea L. J Plant Physiol 141:269–275. https://doi.org/10.1016/S0176-1617(11)81733-5
Seitz HU, Gärtner DE (1994) Enzymes in cardenolide-accumulating shoot cultures of Digitalis purpurea L. Plant Cell Tissue Organ Cult 38:337–344. https://doi.org/10.1007/BF00033894
Gartner DE, Keilholz W, Seitz HU (1994) Purification, characterization and partial peptide microsequencing of progesterone 5β-reductase from shoot cultures of Digitalis purpurea. Eur J Biochem 225:1125–1132. https://doi.org/10.1111/j.1432-1033.1994.1125b.x
Palazón J, Bonfill M, Cusidó RM et al (1995) Effects of auxin and phenobarbital on morphogenesis and production of digitoxin in Digitalis callus. Plant Cell Physiol 36:247–252. https://doi.org/10.1093/oxfordjournals.pcp.a078756
Bonfill M, Palazón J, Cusidó RM et al (1996) Effect of auxin and phenobarbital on the ultrastructure and digitoxin content in Digitalis purpurea tissue culture. Can J Bot 74:378–382. https://doi.org/10.1139/b96-047
Pérez-Alonso N, Chong-Pérez B, Capote A et al (2014) Agrobacterium tumefaciens-mediated genetic transformation of Digitalis purpurea L. plant. Biotechnol Rep 8:387–397. https://doi.org/10.1007/s11816-014-0329-0
Cacho M, Morán M, Herrera MT, Fernández-Tárrago J (1991) Morphogenesis in leaf, hypocotyl and root explants of Digitalis thapsi L. cultured in vitro. Plant Cell Tissue Organ Cult 25:117–123. https://doi.org/10.1007/BF00042182
Sales E, Nebauer SG, Arrillaga I, Segura J (2002) Plant hormones and Agrobacterium tumefaciens strain 82.139 induce efficient plant regeneration in the cardenolide-producing plant Digitalis minor. J Plant Physiol 159:9–16. https://doi.org/10.1078/0176-1617-00534
Sales E, Segura J, Arrillaga I (2003) Agrobacterium tumefaciens-mediated genetic transformation of the cardenolide-producing plant Digitalis minor L. Planta Med. 69: 143–147. https://doi.org/10.1055/s-2003-37709
Kreis W, Haug B, Yücesan B (2015) Somaclonal variation of cardenolide content in Heywood’s foxglove, a source for the antiviral cardenolide glucoevatromonoside, regenerated from permanent shoot culture and callus. In Vitro Cell Dev Biol Plant 51:35–41. https://doi.org/10.1007/s11627-014-9642-0
Pérez-Alonso N, Martín R, Capote A et al (2018) Efficient direct shoot organogenesis, genetic stability and secondary metabolite production of micropropagated Digitalis purpurea L. Ind Crop Prod 116:259–266. https://doi.org/10.1016/j.indcrop.2018.02.067
Pérez-Alonso N, Wilken D, Gerth A et al (2009) Cardiotonic glycosides from biomass of Digitalis purpurea L. cultured in temporary immersion systems. Plant Cell Tissue Organ Cult (PCTOC) 99:151–156. https://doi.org/10.1007/s11240-009-9587-x
Patil JG, Ahire ML, Nitnaware KM et al (2013) In vitro propagation and production of cardiotonic glycosides in shoot cultures of Digitalis purpurea L. by elicitation and precursor feeding. Appl Microbiol Biotechnol 97:2379–2393. https://doi.org/10.1007/s00253-012-4489-y
Li Y, Gao Z, Piao C et al (2014) A stable and efficient Agrobacterium tumefaciens-mediated genetic transformation of the medicinal plant Digitalis purpurea L. Appl Biochem Biotechnol 172:1807–1817. https://doi.org/10.1007/s12010-013-0648-6
Verma SK, Gantait S, Jeong BR, Hwang SJ (2018) Enhanced growth and cardenolides production in Digitalis purpurea under the influence of different LED exposures in the plant factory. Sci Rep 8:18009. https://doi.org/10.1038/s41598-018-36113-9
Martín R, Chong-Pérez B, Pérez-Alonso N (2015) Organogénesis in vitro en el género Digitalis. Biotecnología Vegetal 15:195–206
Bairu MW, Aremu AO, Van Staden J (2011) Somaclonal variation in plants: causes and detection methods. Plant Growth Regul 63:147–173. https://doi.org/10.1007/s10725-010-9554-x
Govindaraj M, Vetriventhan M, Srinivasan M (2015) Importance of genetic diversity assessment in crop plants and its recent advances: an overview of its analytical perspectives. Genet Res Int 2015(14). https://doi.org/10.1155/2015/431487
Gavidia I, Del Castillo Agudo L, Pérez-Bermúdez P (1996) Selection and long-term cultures of high-yielding Digitalis obscura plants: RAPD markers for analysis of genetic stability. Plant Sci 121:197–205. https://doi.org/10.1016/S0168-9452(96)04510-4
Sales E, Nebauer SG, Arrillaga I, Segura J (2001) Cryopreservation of Digitalis obscura selected genotypes by encapsulation-dehydration. Planta Med 67:833–838. https://doi.org/10.1055/s-2001-18861
Nebauer SG, Del Castillo-Agudo L, Segura J (2000) An assessment of genetic relationships within the genus Digitalis based on PCR-generated RAPD markers. Theor Appl Genet 100:1209–1216. https://doi.org/10.1007/s001220051426
Sales E, Nebauer SG, Mus M, Segura J (2001) Population genetic study in the Balearic endemic plant species Digitalis minor (Scrophulariaceae) using RAPD markers. Am J Bot 88:1750–1759. https://doi.org/10.2307/3558349
Boronnikova SV, Kokaeva ZG, Gostimskiǐ SA et al (2007) Analysis of DNA polymorphism in a relict Uralian species, yellow foxglove (Digitalis grandiflora Mill.), using RAPD and ISSR markers. Genetika 43:653–659. https://doi.org/10.1134/S1022795407050080
Hodel RGJ, Segovia-Salcedo MC, Landis JB et al (2016) The report of my death was an exaggeration: a review for researchers using microsatellites in the 21st century. Appl Plant Sci 4:1600025. https://doi.org/10.3732/apps.1600025
Pérez-Alonso N, Chong-Pérez B, Capote A et al (2016) Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants, 2nd edn. Springer, New York
Pérez-Alonso NL, Arana LF, Capote PA et al (2014) Estimulación de cardenólidos en brotes de Digitalis purpurea L. cultivados in vitro mediante elicitores. Rev Colomb Biotecnol 16:51–61
Sales E, Muñoz-Bertomeu J, Arrillaga I, Segura J (2007) Enhancement of Cardenolide and Phytosterol levels by expression of an N-terminally truncated 3-Hydroxy-3-methylglutaryl CoA Reductase in transgenic Digitalis minor. Planta Med 73:605–610. https://doi.org/10.1055/s-2007-967199
Kairuz E, Pérez-Alonso N, Chong-pérez B (2018) Estrategias para la selección in vitro de plantas transgénicas de Digitalis L. Biotecnologia Vegetal 18(2):63–80
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Nature Switzerland AG
About this entry
Cite this entry
Kairuz, E., Pérez-Alonso, N., Angenon, G., Jiménez, E., Chong-Pérez, B. (2021). Shoot Organogenesis, Genetic Stability, and Secondary Metabolite Production of Micropropagated Digitalis purpurea. In: Ramawat, K.G., Ekiert, H.M., Goyal, S. (eds) Plant Cell and Tissue Differentiation and Secondary Metabolites. Reference Series in Phytochemistry. Springer, Cham. https://doi.org/10.1007/978-3-030-30185-9_16
Download citation
DOI: https://doi.org/10.1007/978-3-030-30185-9_16
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-030-30184-2
Online ISBN: 978-3-030-30185-9
eBook Packages: Chemistry and Materials ScienceReference Module Physical and Materials ScienceReference Module Chemistry, Materials and Physics